The heavy metal cadmium is a xenobiotic toxicant of environmental and occupational concern and it has been classified as a human carcinogen. Inhalation of cadmium has been implicated in the development of emphysema and pulmonary fibrosis, but, the detailed mechanism by which cadmium induces adverse biological effects is not yet known. Therefore, we undertook the investigation of genes that are induced after cadmium exposure to illustrate the mechanism of cadmium toxicity For this purpose, we employed the polymerase chain reaction-based suppression subtractive hybridization technique. We identified 29 different cadmium-inducible genes in human peripheral mononuclear cells, such as macrophage migration inhibitory factor, lysophosphatidic acid acyltransferase-${\alpha}$, enolase-1${\alpha}$, VEGF, Bax, neuron-derived orphan receptor-1, and Nur77, which are known to be associated with inflammation, cell survival, and apoptosis. Induction of these genes by cadmium treatment was further confirmed by semi-quantitative reverse-transcription polymerase chain reaction. Further, we found that these genes were also induced after cadmium exposure in normal human lung fibroblast cell line, WI-38, suggesting potential use of this induction profile to monitor cadmium toxicity in the lung. Next, Nur77, one of cadmium-inducible genes, was further studied since the products of Nur77 are known to be involved in the apoptotic process of lung cells. Following cadmium treatment, Nur77 gene expression was increased at protein-level in A549 cells. Consistently, the reporter containing Nur77 binding sequence was activated by 2.5-fold after exposure to cadmium in reporter gene analysis by transient transfection experiments. When the plasmid encoding dominant negative Nur77 that represses the transcriptional function of wild-type Nur77 was transfected into A549 cells, the expression of Bax was significantly reduced, suggesting that induction of Nur77 was an important process in cadmium-induced apoptosis in the cells. Cadmium induced the expression of Nur77 in vivo, confirming the relevance of the data obtained in viro. Together our results suggest that Nur77 gene expression in exposure to cadmium leads apoptosis of lung cells which may cause pathological changes in lung.

A method for the determination of alkylphenols, chlorophenols and bisphenol A in paper materials using GC/MS-SIM has been developed. Eleven endocrine disrupting chemicals (EDCs) of phenols in paper samples were extracted with acetonitrile. Also, solid-phase extraction (SPE) with XAD-4 and subsequent conversion to isobutoxycarbonyl derivatives or tert.-butyldimethylsilyl derivatives for sensitive analysis with the selected ion-monitoring (SIM) mode. The recoveries were 82.4∼108.8 % by area ratio of pheranthrene-d$\sub$10/ vs bisphenol A d$\sub$l6/. (isoBOC derivatization and TBDMS derivatization) The SIM responses were linear with the correlation coefficient varying 0.9717∼0.9995 (isoBOC derivatization), and 0.9842∼0.9980 (TBDMS derivatization). The range of concentrations was respectively, 0.95∼l.44 ng/g in 2,4-dichlorophenol, 1.01∼1.17 ng/g in t-butylphenol, 2.17∼5.84 ng/g in pentachlorophenol, 12.68∼14.88 ng/g in nonylphenol and 30.84∼153.72 ng/g in bisphenol A.

The free-living nematode, Caenorhabditis elegans (C. elegans) has been adopted as a multicellular biosensor of biological toxicity for alkylphenol, organotin compounds and heavy metals. To adopt as a biosensor, suitability to assess must be fulfilledthrough several criteria; the organism must be sensitive to the testing toxicants, easy to manage in the laboratory and available throughout the year. C. elegans widely used as a simple multicellular organism in developmental biology studies and satisfies all these criteria, and its culture conditions, developmental staging, anatomy and genetic properties are well defined. In addition, researchers can take advantage of the worm's short life cycle, low cost and little individual variation. Moreover, genomic sequencing of C. elegans has recently been completed. With these aspectsof the organism, C. elegans become a more potent model organism for basic and applied bioassays.

This study is to investigate the environmental levels and trend of dioxins, which was the 3$\^$rd/ year of environmental monitoring research for endocrine disrupting chemicals since 1999. Total 282 samples were analyzed from 115 sites including 26 sites of airs, 43 sites of waters, 11 sites of sediments and 35 sites of soil, which were the same as those of investigated sites in 2000. Sampling period was from June 2001 to June 2002. Target chemicals were seventeen species of 2,3,7,8-chlorine-substituted PCDD and PCDF congeners and were analyzed by the standard methods, established by National Institute Environmental Research (NIER). The average concentration of dioxins in air decreased from 0.324 pg-TEQ/N㎥ in 2000 to 0.287 pg-TEQ/N㎥ in 2001, and those in water and soil were 0.073pg-TEQ/L and 1.703pg-TEQ/dry g, respectively, which was the less values detected in 2000. In sediment, however, the value was 0.086pg-TEQ/dry g, which was the increase from the value of the year 2000. The concentration range of dioxins in air for 26 sites in 17 regions detected were 0.013∼l.664pg-TEQ/N㎥, 4 sites from those were exceeded the Air Quality Standards of Dioxin in Japan (0.6 pg-TEQ/N㎥). The tolerable daily intake of dioxins was calculated at the highest level (1.664) in air, with referring the soil and food data from Japan, was calculated to be 2.85pg-TEQ/kg/day, which was below the level of 4 pg-TEQ/kg/day suggested in KFDA(Korea). While the average concentration of dioxins in 15 big cities was 0.190 pg-TEQ/N㎥, that in 8 medium/small cities constituting an industrial complex was 0.558 pg-TEQ/N㎥. In water, the concentration range detected were 0∼0.946pg-TEQ/L and the trend of the average concentrations shows an increase from those of 1999 but decreased from those of 2000, any sites however were not exceeded the Water Quality Standards of Dioxin in Japan (1 pg- TEQ/L). In soil. the detected range were 0∼43.333 pg-TEQ/dry g and the average concentration decreased, compared with the results of 2000. According to the monitoring results by land utilization, the detected range were 0∼43.333pg-TEQ/dry g in farmland, 0.017∼0.601 pg-TEQ/dry g in the industrial area, 0.005∼0.049pg-TEQ/dry g in the park and 0.008∼1.825 pg-TEQ/dry g in the rest. In sediment, the detected range increased from 0∼0.244 pg-TEQ/dry g to 0∼0.537 pg-TEQ/dry g, based on the results of 2000. For the proper control of dioxins, continuous monitoring needs to be performed and in addition, the dioxin inventory should be prepared for major sources through the dioxin emission survey. These results would provide sound and solid basis for proper decision making of dioxins management like establishment of environmental quality standards in Korea.

To understand environmental paths of the transport and accumulation of endocrine disrupting chemicals (EDCs), a single cell multimedia fate model has been constructed and evaluated. The EDCs of concern were PAHs, Organochlorine Pesticides (OCPs), PCBs, Alkyl phenols, and phthalates. An evaluation model was designed for the multimedia distribution, including air, water, soil, sediment and vegetation. This model was verified using reported values and via monitoring data. Based on collected data, the distribution trends of EDCs with respect to environmental media were analyzed. Those results have applied to the model for the prediction of the spatial and temporal distribution of EDCs in Seoul. Especially, phenol compound, phthalates, PAHs, PCBs and organochlorine pesticides were estimated and the model was verified. This model was successfully conducted to environmental media, such as air (vapor and suspended particles), soils (forest soil, bare soil, and cement-concrete covered soil), water (dissolved and suspended solids), sediment, trees (deciduous and coniferous). The discrepancies between the model prediction and the measured data are approximately within or near a factor of 10 for the PAHs of three rings through that of six rings, implying that multimedia distribution of the PAHs could be predicted with a factor of 10. Concerning about the air equilibrium may be assumed, a fugacity at steady state is similar in all environmental media. Considering the uncertainties of this model, the use of equilibrium models may be sufficient for assessing chemical fates. In this study, a suggestion was made that modeling and estimation of chemicals in environmental multimedia be rigorously evaluated using the measured flux data. In addition, these data should be obtained, for example, from the precise and standardized inventory of the target chemicals. The model (EDC Seoul) will be refined in an on-going research effort and will be used to support decision-making concerning the management of EDCs.

Benzoyl peroxide is a high production volume chemical, which was produced about 1,375 tons/year in Korea as of 2001 survey. Most of them are used as initiators in polymerization, catalysts in the plastics industry, bleaching agents for flour and medication for acne vulgaris. The substance is one of the sever chemicals of which human and environmental risks are being assessed by National Institute of Environmental Research under the frame of OECD SIDS Program. It has a melting point of 104-106 $^{\circ}C$ and has solubility of 9.1 mg/1 in water at 25 $^{\circ}C$. The substance was readily biodegradable (83 % after 21days) and had toxic effects to aquatic organisms. The range of 72 hr-EbC50 (biomass) for algae was 0.07-0.44 mg/1 and 48 hr-EC50 for daphnia was 0.07-2.91 mg/1. The LC50 of acute toxicity to fish was 0.24-2.0 mg/1. Although the toxic effects of benzoyl peroxide to aquatic organisms were investigated, environmental monitoring data were not studied. In this study, distribution of the chemical among multimedia environment was estimated using EQC model based on the physical-chemical properties to evaluate the risk of benzoyl peroxide in environment. In level I, II calculation the chemical was distributed to soil (68.3 %) and water (28.7 %). In level III calculation it was primarily distributed to soil (99.9 %) and overall residence time of 3.4 years was estimated. Benzoyl peroxide could be persistent in environment.

The goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals. Japanese medaka was treated under appropriate sublethal concentrations of copper and the fish were subjected to copper treatment after starvation for 48 hr. The untreated individuals showed common behavioral characteristics (i.e., smooth and linear movements with small curvatures) in the movement behaviors.

Fluoranthene, a common polycyclicaromatic hydrocarbon (PAH), exhibits phototoxicity which may affect aquatic organisms. The eventual goal of this study is to develop a biomarker used in monitoring abnormal behaviors of Japanese medaka (Oryzias latipes) as a model organism caused by hazardous chemicals that are toxic and persistent in the ecosystem. In this study, we investigated neural toxicity of fluoranthene in Japanese medaka (Oryzias latipes) which was correlated with its behaviors.

Plasma melatonin in the seasonal light-period is circadian rhythmically secreted. Maximal secretion showed at 14 o'clock in summer- and winter-like period, but minimal secretion showed at 5 o'clock in summer-like period and at 8 in winter-like period. These times of minimal secretions were at the beginning of light period. Plasma melatonin in the light period is secreted 62.5% more than in the dark period in summer-like period and 45.9% more in winter-like period. Total plasma melatonin in winter-like period is secreted 56.5% more than in summer-like period. The weights of testis(-20.8%) and body(-7.1%) were reduced in the winter-like period. By the increase of plasma melatonin in mice, body- and testis-weights are decreased, on the contrary by the decrease of plasma melatonin in mice, body and testis weights are increased. In view of the results so far achieved melatonin changes on the body weight and reproductive organ in mice. It is presumed that melatonin effects on the metabolism and sex hormone.

전국 주요 산단 지역에서 현재 유통되고 있는 화학물질의 스크리닝 수준에서의 생태 위해성 평가를 수행하고, 체계적인 위해도 분석 시스템을 구축하기 위한 수생 및 육상 생태계가 통합된 생태 위해성 예측 모형을 개발하였다. 전국 118개 공단 중 시화반월, 전주, 대전, 대구, 청주, 울산, 구미, 여천공단 등 8개 주요 산단 지역을 program 개발을 위한 비교 대상 지역으로 선정하였고 이들 공단에서 사용되고 있는 약 1700여종의 화학물질의 물리 화학적 특성 자료와 생태 독성값의 문헌치, 추정치를 포함한 database를 구축하였다. 구축된 database를 활용하여 산단별, 화학물질별 생태 위해도를 상대적으로 비교한 결과 어류 만성의 위해도는 청주 공단>울산미포>여천 공단의 순이었고, 물벼룩 만성의 위해도는 시화반월>여천>울산미포 공단의 순이었다. 또한 산단별로 위해도 우선순위 물질을 선정하였으며 styrene, xylene 등의 위해도가 높게 나타났다. 이때 화학 물질의 수계 배출량은 TRI 자료와 사용량과의 상관관계로부터 구한 배출 계수를 적용하여 산정하였으며, 선정된 위해도 우선 순위 물질을 대상으로 실제 공단의 하천 시료를 채취, 분석하여 이 모형에 의해 예측된 생태 위해도의 타당성을 검증하고 보완하는 연구를 지속적으로 수행하여 예측 program의 신뢰도를 높이고자 한다.

산업의 발달정도, 업종, 규모 등에 따라 수역으로 배출되는 특정수질유해물질은 해마다 그 수, 양, 독성이 증가하여 인간과 생태계에 미치는 영향이 매우 커서 이를 예측하고 관리해야하는 필요성이 점차 증대되고 있다. 이에 특정수질유해물질과 관련된 산업폐수관리법 및 수질기준법의 국가별 현황을 조사하여보고, 궁극적인 수 환경보전목표와의 연관성을 검토해봄으로써, 국내 특정수질유해물질관리의 문제점을 확인한 후에 배출시설관리체계 개선을 위한 국내 배출허용기준을 제안해 보고자 한다.

In terms of the risk assessment, qualitative and quantitative informations are needed to estimate the exposures of environmental pollutants, which may be potentiality of risks, and those are the information about the changes caused by the chemical transportation among environmental media and transformation in environmental media by duration. The various fate mechanism of chemical is possible for estimation of chemical concentration in environmental media. Since there are limitations in measuring the change of chemical concentration within all medium according to the time period, estimating method through modeling are developed.

In the previous studies, some genetic polymorphisms in the human mitochondrial DNA have been associated with athletic performance in several populations. To investigate the relationship between mitochondrial DNA 5178A/C polymorphism and athletic performance in Korean population, blood samples were collected from 100 male Korean elite athletes and 64 sedentary controls. There was no significant difference in allele frequency of mitochondrial DNA 5178A/C polymorphism between two groups (P > 0.05). However, 5178A allele frequency in Korean population was very higher than those in other populations studied. Because it has been reported that this genetic polymorphisms is associated with longevity, further study will be needed to clarify the relationship between this genetic polymorphism and life expectancy of Korean population.

Adult periodontitis is a chronic inflammatory disease whose etiology is not well defined. Recent studies have shown that vitamin D receptor gene has been a candidate for the susceptibility of adult periodontitis. The purpose of this study is to investigate the frequency of Taq I restriction fragment length polymorphism (RELP) in the vitamin D receptor gene in 141 periodontically healthy controls and 32 adult periodontitis patients. Taq I RFLP in the vitamin D receptor gene were detected by PCR amplification, followed by restriction enzyme digestion and 2% agarose gel electrophoresis. There were no significant difference in the distribution of Taq I RFLP between healthy controls and adult periodontitis group (P > 0.05). Thus, Taq I RFLP in the vitamin D receptor gene may not confer the susceptibility to adult periodontitis in Korean population. However, t allele distributions of this RFLP showed various frequencies among ethnic groups studied. Further studies in other ethnic groups will be required.

The testis is composed of four cell types like supporting cells, steroid-producing cells, connective tissue cells and germ cells. Apoptosis is a common phenomenon during spormatogenesis. Apoptosis of germ cells can also be induced by exposure to radiation. Previous studies have shown that most types of germ cells are rather radiosensitive while somatic cells in testis are much more radio-resistant. The somatic cells in testis are divided to mainly Sertoli and Leydig cells. Though somatic cells are more radio-resistant than germ cells, radiation can induce the impairment of their function. This damaged function of somatic cells may accelerates degeneration of germ cell indirectly. Tn the present study, we have examined the apoptotic effect of mouse testis and irradiation effect of steroidogenesis of Leydig cells after irradiation.

Transcript profiling is a particularly valuable tool in the field of steroid receptor biology, as these receptors are ligand-activated transcription factors and therefore exert their initial effects through altering gene expression in responsive cells. Also, an increased awareness of endocrine disrupting chemicals (EBCs) and their potential to affect wildlife and humans has produced a demand for practical screening methods to identify endocrine activity. Here we developed an in-house cDNA microarray, named KISTCHIP-400, with 401 clones, hormone related genes, factors, and ESTs, based on public database and research papers. Theses clones contained estrogen, androgen, thyroid hormone St receptors, sex hormone signal transduction ＆ regulation, c-fos, c-myc, ps2 gene, metabolism related genes etc. And to validate the KISTCHIP-400, we investigated gene expression profiles with reference hormones, 10$\^$-8/ M 17be1a-estradiol, 10$\^$-7/ M testosterone, 10$\^$-7/ M progesterone, and thyroxin in MCF-7 cell line. Although it is in first step of validation, low doses and combinations of EDCs need to be tested. Our preliminary results that indicate the developed microarray may be a useful laboratory tool for screening EDCs and elucidating endocrine disrupting mechanism.

Exposure to some synthetic environmental chemicals and their metabolites cause reproductive problems in a variety of vertebrate via endocrine mechanisms. However, in most cases, the link between these compounds and adverse effects on humans, fish, and wildlife has not been established, which necessitates a closer look at the molecular, functional, and clinical implications of these chemicals in the environment. Calbindin-D9k (CaBP-9k) is a member of a largo family of intracellular calcium binding proteins that have high affinities for calcium. It was reported that the estrogen level of uterus affected the expression of the CaBP-9k gene in rat uterus. We examined the dose-dependent CaBP-9K gene expression in the uterus for three-days injection of methoxychlor (HC) in the overectomized immature rats and the relation with uterotrophic response of the compoundsand compared the responses induced by MC according to the route of administration.

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). It has been reported to have an anti-inflammatory effect on rat paw edema model. To develop as an anti-allergic drug, genotoxicity of sophoricoside was investigated in bacterial and mammalian cell system such as Ames bacterial test, chromosomal aberration assay, Comet assay and MOLY assay. In Ames test, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/plate concentrations was not shown significant mutagenic effect in Salmonella typhimurium TA98, TA100, TA1535 and TA1537 strains. The cytotoxicity (IC$\_$50/ and IC$\_$20/) of sophoricoside was determined above the concentration of 5000 $\mu\textrm{g}$/ml in Chinese hamster lung (CHL) fibroblast cell and L5178Y mouse lymphoma cell line. At concentrations of 5000, 2500 and 1250 $\mu\textrm{g}$/ml, this compound was not induced chromosomal aberration in CHL fibroblast cell in the absence and presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in L5178Y cell line. Also in MOLY assay, sophoricoside of 5000 ∼ 313 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in absence of S-9 metabolic activation system. However, the higher concentration of 5000 and 2500 $\mu\textrm{g}$/ml of sophoricoside induced the increased mutation frequency (MF) in the presence of S-9 metabolic activation system. From these results, no genotoxic effects of sophoricoside observed in bacterial systems whereas, genotoxic effects observed in mammalian cell systems in the presence of metabolic activation system. These results suggested that the metabolite(s) of sophoricoside can cause some genotoxic effects in mammalian cells.

To develope the novel anti-allergic drug, many sophoricoside derivatives were synthesized. Among these derivatives, JSH-II-3, JSH-Ⅵ-3, JSH-Ⅶ-3, and JSH-Ⅷ-3 were selected and subjected to high throughput toxicity screening (HTTS) because they revealed strong IL-5 inhibitory activity and limitation of quantity. Mouse lymphoma thymidine kinase (tk$\^$+/-/) gene assay (MOLY) and single cell gel electrophoresis (Comet) assay in mammalian cells were used as HTTS tool in our laboratory. In MOLY assay, JSH-Ⅶ-3 at 50 ∼ 6 $\mu\textrm{g}$/ml concentrations was not shown significant mutagenic effect in the absence and presence of S-9 metabolic activation system. However, the concentration of ISH-II-3, 38 $\mu\textrm{g}$/ml, induced increased mutation frequency (MF) in the presence of S-9 metabolic activation system. Also in comet assay, DNA damage was not observed in JSH-Ⅵ-3 and JSH-Ⅶ-3, wherase concentration of 32.8 $\mu\textrm{g}$/ml in JSH-II-3 and 13.9 $\mu\textrm{g}$/ml in JSH-Ⅶ-3 were induced DNA damage in the absence of S-9 metabolic activation system. Therefore, we suggest that JSH-Ⅵ-3 and JSH-Ⅶ-3 have no genotoxic effects but JSH-II-3 and JSH-Ⅷ-3 induce some mutagenicity and DNA strand breaks in mouse lymphoma cell line used this study.

CKD-712, named S-YS49 is a chiral compound derived from higenamine (one component of Aconite spp.) derivatives. To compare the cytotoxicity of CKD-712 between in the absence and in the presence of S9 metabolic activation system, we performed trypan blue dye exclusion assay in Chinese hamster lung (CHL) cell. In CHL cells, the cytotoxicity (IC50) of CKD-712 was 92.9 $\mu\textrm{g}$/ml and 186.1 $\mu\textrm{g}$/ml in the absence and presence of S9 metabolic activation, respectively. And we also investigated the induction of DNA damages in mammalian cells. To perform the single cell gel electrophoresis, we determined optimum concentration in mouse lymphoma L5178Y cells using frypan blue dye exclusion assay Each IC20 of CKD-712 was determined the concentration of 23.4 $\mu\textrm{g}$/ml and 24.8 $\mu\textrm{g}$/ml in the absence and presence of S9 metabolic activation, respectively. In the comet assay, DNA damage was not observed at the concentration range from 23.4 $\mu\textrm{g}$/ml to 5.9 $\mu\textrm{g}$/ml in the absence of S9 metabolic activation system. In the presence of S9 metabolic activation system, DNA damage was not observed at the concentration range from 24.8 $\mu\textrm{g}$/ml to 6.2 $\mu\textrm{g}$/ml. From these results, it is assumed that CKD-712 may be metabolized to less cytotoxic metabolite(s).

Methylmercury (MeHg), one of the heavy metal compounds, can cause severe damage to the central nervous system in humans. Many reports have shown that MeHg is poisonous to human body through contaminated foods and has released into the environment. Despite many studies on the pathogenesis of MeHg-induced central neuropathy, no useful mechanism of toxicity has been established so far. In this study, two methods, cDNA Microarray and SSH, were performed to assess the expression profile against MeHg and to identify differentially expressed genes by MeHg in neuroblastoma cell line. TwinChip Human-8K (Digital Genomics) was used with total RNA from SH-SY5Y (human neuroblastoma cell line) treated with solvent (DMSO) and 6.25 uM (IC50) MeHg. And we performed forward and reverse SSH method on mRNA derived from SH-SY5Y treated with DMSO and MeHg (6.25 uM). Differentially expressed cDNA clones were sequenced and were screened by dot blot and ribonuclease protection assay to confirm that individual clones indeed represent differentially expressed genes. These sequences were identified by BLAST homology search to known genes or expressed sequence tags (ESTs). Analysis of these sequences may provide an insight into the biological effects of MeHg in the pathogenesis of neurodegenerative disease and a possibility to develop more efficient and exact monitoring system of heavy metals as environmental pollutants.

Examination was made of the urinary metabolite(s) of CKD-712, which is a chiral compound, named S-YS49 derived from higenamine (one component of Aconite spp.) derivatives. First of all, to analyze the metabolite(s) of CKD-712, a simple and sensitive detection method for CKD-712 was developed by using gas chromatography-mass spectrometry GC/MS). Urine was collected from adult male Sprague-Dawley rats 250${\pm}$10g) in metabolic cage for 24hr after oral administration of 100 mg/kg of CKD-712. The recovery of CKD-712 after extraction and concentration with AD-2 resin column was above 90 % from rat urine. The detection limits of CKD-712 in urine was approximately 0.1 ng/mL. It has well been suggested that isoquinoline possessing catechol moiety such as CKD-712 should be subjected to the catechol-O-methyl kransferase activity in vivo. We detected three major peaks of presumed CKD-712 metabolites in the total ion chromatogram obtained from the rat urine sample after oral administration of CKD-712. From these results, it is assumed that the urinary metabolites are mono-methylation in the naphthyl moiety (metabolite I ), methylation at the C-6 or 7 hydroxy group in the isoquinoline moiety and hydroxylation at in the naphthyl moiety (metaboliteII), and methylation at the C-6 or 7 hydroxy group in the isoquinoline moiety (metaboliteIII).

Sophoricoside was isolated as the inhibitor of IL-5 bioactivity from Sophora japonica (Leguminosae). To develope as novel anti-allergic drug, kinetic study was performed in rats. Serum concentration of sophoricoside was measured by gas chromatography-mass spectrometry (GC/MS) in male Sprague-Dawley rat (250${\pm}$10g, n=5) after oral administration of sophoricoside (100mg/kg). The recovery of sophoricoside after extraction and concentration was above 95 % from rat serum. Between-day precision(relative standard deviation 2.2-2.8%) and within-day precision(2.0-12.1%) were determined from replicate analysis of a spiked control and incurred serum sample. The detection limits of sophoricoside in this serum was approximately 0.1 ng/mL. The Pharmacokinetic parameters were derived from the noncompartmental analysis. The C$\_$max/(3.56${\pm}$0.34 $\mu\textrm{g}$/mL) value for sophoricoside in male rat was observed at 7.6 h. The elimination half-life(t$\_$1/2/) of sophoricoside was approximately 4.47 h, the mean residence time (MRT) averaged 10.75 h, the total body clearance (Cl) averaged 0.0042 mL/min/kg. and the area under the serum concentration-time curve (AUC$\_$0-$\infty$/) was 24.93 $\mu\textrm{g}$$.$hr/mL.

Endometriosis is classically defined as the growth of endometrial glands and stroma at extrauterine sites. Although it is a common gynecological problem accompanied by chronic pelvic pain, infertility, and adhesion formation, the etiology of this disease is unknown. Endometriosis pathogenesis may involve endocrine and immune dysfunction since uterine endometrial growth is regulated by sex hormones in concert with bioactive mediators produced by uterine immune and endocrine cells. Thus, exposure to environmental toxicants disrupting endocrine and immune responses potentially affect the development and progression of endometriosis.

Pomegranate, a small tree originating in Orient, belongs to Punicaceae family. The seeds contain an oil of which about 80% is rare trans 18 carbon fatty acid (punicic acid), and have highest botanical concentration of a sex steroid, estrone. Pharmacological properties of pomegranate extract have been studied, with anti-microbial, anti-parasitic, ant-viral, and anti-cancer effects. We have examined the estrogenic activity of the pomegranate extracts using MCP-7-ERE cells. MCF-7-ERE cells, stably transfected with pERE-Luc were treated various amount of pomegranate extract and after overnight treatments, luciferase activity were measured. Estradiol(E2) dose dependently induced luciferase activity in this cell and maximal response is obtained at 100pM E2.

Recent industrial society has human widely exposed to PAHs that are coming from the incomplete combustion of organic material as widespread environmental contaminants. Biological activities of PAHs are not known although PAHs are considered as carcinogens. The mechanism of action of PAHs has been studied extensively, however it is not clear how PAHs turn on CYPlAl in human breast cancer, Our laboratory have been studied the effect of PAHs in the human breast cancer cells, MCF-7. In this study, we examined the ZR-75-1, human breast cancer cells, as a new system to evaluate bioactivity of PAHs and to compare the PAHs action with that of MCF-7 cells. ZR-75-1 human breast cancer cell line is responsible to estrogen and progesterone. We have been able to establish long term culture system of this cells then used for the study to the effect of 13 different PAHs and environmental samples. We demonstrate that PAHs induced the CYP1A1 promoter and 7-ethoxyresorufin O-deethylase (EROD) activity in a concentration-dependent manner. RT-PCR analysis indicated that PAHs significantly up-regulate the level of CYP1A1 mRNA. Some of PAHs showed stronger stimulatory effect on CYP1 gene expression than TCDD Apparently, ZR-75-1 cells have Aryl hydrocarbon receptors (AhR), therefore it would be a good experimental tool to study the cross-talk between PAHs and steroid actions.

The International Agency for Research on Cancer (IARC, 1989) has classified whole diesel exhaust as probably carcinogenic to humans. Diesel exhaust particulate matter (DPM) adsorbs different chemical substances including PAHs and nitroarenes. DPM is emphasized because it is a major component of diesel exhaust, it is suspected of contributing to a health hazard. Diesel exhaust is a complex mixture of carbon particles and associated organics and inorganics, and it is not known what fraction or combination of fractions cause the health effects [cancer effects, noncancer effects (respiratory tract irritation/inflammation and changes in lung function)] that have been observed with exposure to diesel exhaust. In order to identify which chemical classes are responsible for the majority of the observed biological activities, we performed a particular biological/chemical analysis. Respirable particulate matter (PM2.5: <2.5mm) was collected from diesel engine exhaust using a high-volume sampler equipped with a cascade impactor. Particulate oganic matter was extracted by the dichloromethane/sonication method and the crude extract was fractionated according to EPA recommended procedure into seven fractions by acid-base partitioning and silica gel column chromatography. We examined genotoxic potentials of diesel exhaust particulate matter using novel genotoxicity tests, which are rapid, simple and sensitive methods for assessing DNA-damage at the DNA and chromosomal level (comet assay, in vitro MN test and Ames test). Higher genotoxic potency was observed in non polar fractions and several PAHs were detected by GC-MS, such as 1,2,5,6 dibenzanthracene, chrysene, 1,2-benzanthracene, phenanthrene and fluoranthene.

It is well known that diesel exhaust particulate matter contains mutagenic PAHs, such as benzo[${\alpha}$]pyrene, benz[${\alpha}$]anthracene, chrysene, etc. Therefore it is suspected that these chemicals act on estrogen receptor and reveal endocrine-disrupting effects. Recent attention has focused on causative chemicals of endocrine-disrupting effects. We examined the estrogenic activity of respirable diesel exhaust particulate matter derived from diesel powered vehicle. PM2.5 diesel exhaust of vehicle was collected using a high volume sampler equipped with a cascade impactor. Diesel exhaust samples were fractionated according to EPA methods. The presence of estrogenic and antiestrogenic chemicals in PM 2.5 diesel exhaust was determined using E-screen assay. To quantitatively assess the estrogenic and antiestrogenic activities in diesel exhaust particulate matter, estradiol equivalent concentration (bio-EEQ) was calculated by comparing the concentration response curve of the sample with those of the estrogen calibration curve. Weak estrogenic activities and strong antiestrogenic activities were detected in the crude extract and moderately polar fractions. Higher antiestrogenic potency was observed with higher EROD activities in aliphatic and aromatic compounds fraction. In conclusion, estrogenic/antiestrogenic-like activities were present in diesel exhaust particulate matter. However, the health consequences of this observation was unknown, the presence of these activities may contribute to and exacerbate adverse health effect evoked by diesel exhaust particulate matter.